Affiliation:
1. Diagnostic and Reference Laboratory, Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado
Abstract
ABSTRACT
The dengue (DEN) viruses are positive-strand RNA viruses in the genus
Flavivirus
. Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are important human arboviral diseases caused by infection with one of four closely related but serologically distinct DEN viruses, designated DEN-1, DEN-2, DEN-3, and DEN-4 viruses. All four DEN serotypes are currently cocirculating throughout the subtropics and tropics, and genotypic variation occurs among isolates within a serotype. A real-time quantitative nucleic acid amplification assay has been developed to detect viral RNA of a single DEN virus serotype. Each primer-probe set is DEN serotype specific, yet detects all genotypes in a panel of 7 to 10 representative isolates of a serotype. In single reactions and in fourplex reactions (containing four primer-probe sets in a single reaction mixture), standard dilutions of virus equivalent to 0.002 PFU of DEN-2, DEN-3, and DEN-4 viruses were detected; the limit of detection of DEN-1 virus was 0.5 equivalent PFU. Singleplex and fourplex reactions were evaluated in a panel of 40 viremic serum specimens with 10 specimens per serotype, containing 0.002 to 6,000 equivalent PFU/reaction (0.4 to 1.2 × 10
6
PFU/ml). Viral RNA was detected in all viremic serum specimens in singleplex and fourplex reactions. Thus, this serotype-specific, fourplex real-time reverse transcriptase PCR nucleic acid detection assay can be used as a method for differential diagnosis of a specific DEN serotype in viremic dengue patients and as a tool for rapid identification and serotyping of DEN virus isolates.
Publisher
American Society for Microbiology
Reference34 articles.
1. Development and Evaluation of Serotype- and Group-Specific Fluorogenic Reverse Transcriptase PCR (TaqMan) Assays for Dengue Virus
2. Chu, M. C., E. J. O'Rourke, and D. W. Trent. 1989. Genetic relatedness among structural protein genes of dengue 1 virus strains. J. Gen. Virol.70:1701-1712.
3. Chungue, E., O. Cassar, M. T. Drouet, M. G. Guzman, M. Laille, L. Rosen, and V. Deubel. 1995. Molecular epidemiology of dengue-1 and dengue-4 viruses. J. Gen. Virol.76:1877-1884.
4. Division of Disease Prevention and Control, Communicable Diseases Program, HCP/HCT, Pan American Health Organization. 1997. Re-emergence of dengue in the Americas. Epidemiolog. Bull.18:1-6.
5. Ferreira, I. B., L. E. Pereira, I. M. Rocco, A. T. Marti, L. T. de Souza, and L. B. Iversson. 1994. Surveillance of arbovirus infections in the Atlantic Forest region, State of Sao Paulo, Brazil. I. Detection of hemagglutination-inhibiting antibodies in wild birds between 1978 and 1990. Rev. Inst. Med. Trop. Sao Paulo36:265-274.
Cited by
356 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献